Abstract
A sequential procedure for single and multiparameter flow cytometry (FCM) that allows detailed cell proliferation and DNA ploidy studies of human solid tumours is reported here. This description includes time collection and storage, tissue disaggregation and staining as well as FCM analysis of samples. The overall feasibility, together with some critical aspects of the DNA/bromodeoxyuridine (BrdU) in vivo assay for cell kinetic studies in human solid tumours, are reported, based on our experience in recent years. We found that the BrdU in vivo method, coupled with bivariate FCM for measurements, is a valuable approach for a 'dynamic' assessment of tumour cell proliferation in the clinical setting. Routine measurements can be achieved providing that well standardised tissue disaggregation and immunolabelling procedures are performed. In different solid tumours, cytokinetic results were satisfactory in 85% of cases as far as the BrdU-labelling index is concerned while 70% were acceptable for DNA synthesis time and tumour potential doubling time. Causes of failure are also considered and discussed. In order to guarantee the finest detection of aneuploidy, routine high resolution single-parameter DNA analysis is suggested. Our experience with further improvements in cell kinetic studies based on the possibility of gating the BrdU/DNA analysis for a subpopulation of interest (i.e. cytokeratin-positive cells in epithelial tumours) is also reported.
| Original language | English |
|---|---|
| Pages (from-to) | 101-113 |
| Number of pages | 13 |
| Journal | Analytical Cellular Pathology |
| Volume | 10 |
| Issue number | 2 |
| Publication status | Published - Mar 1996 |
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Keywords
- DNA-BrdU-cytokeratin
- Flow cytometry
- Proliferation
- Solid tumours
ASJC Scopus subject areas
- Pathology and Forensic Medicine
- Cell Biology
Cite this
Cell proliferation and ploidy of human solid tumours : Methodological experience with in vivo bromodeoxyuridine and DNA flow cytometry. / Mazzini, G.; Danova, M.; Ferrari, C.; Giordano, M.; Dionigi, P.; Riccardi, A.
In: Analytical Cellular Pathology, Vol. 10, No. 2, 03.1996, p. 101-113.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Cell proliferation and ploidy of human solid tumours
T2 - Methodological experience with in vivo bromodeoxyuridine and DNA flow cytometry
AU - Mazzini, G.
AU - Danova, M.
AU - Ferrari, C.
AU - Giordano, M.
AU - Dionigi, P.
AU - Riccardi, A.
PY - 1996/3
Y1 - 1996/3
N2 - A sequential procedure for single and multiparameter flow cytometry (FCM) that allows detailed cell proliferation and DNA ploidy studies of human solid tumours is reported here. This description includes time collection and storage, tissue disaggregation and staining as well as FCM analysis of samples. The overall feasibility, together with some critical aspects of the DNA/bromodeoxyuridine (BrdU) in vivo assay for cell kinetic studies in human solid tumours, are reported, based on our experience in recent years. We found that the BrdU in vivo method, coupled with bivariate FCM for measurements, is a valuable approach for a 'dynamic' assessment of tumour cell proliferation in the clinical setting. Routine measurements can be achieved providing that well standardised tissue disaggregation and immunolabelling procedures are performed. In different solid tumours, cytokinetic results were satisfactory in 85% of cases as far as the BrdU-labelling index is concerned while 70% were acceptable for DNA synthesis time and tumour potential doubling time. Causes of failure are also considered and discussed. In order to guarantee the finest detection of aneuploidy, routine high resolution single-parameter DNA analysis is suggested. Our experience with further improvements in cell kinetic studies based on the possibility of gating the BrdU/DNA analysis for a subpopulation of interest (i.e. cytokeratin-positive cells in epithelial tumours) is also reported.
AB - A sequential procedure for single and multiparameter flow cytometry (FCM) that allows detailed cell proliferation and DNA ploidy studies of human solid tumours is reported here. This description includes time collection and storage, tissue disaggregation and staining as well as FCM analysis of samples. The overall feasibility, together with some critical aspects of the DNA/bromodeoxyuridine (BrdU) in vivo assay for cell kinetic studies in human solid tumours, are reported, based on our experience in recent years. We found that the BrdU in vivo method, coupled with bivariate FCM for measurements, is a valuable approach for a 'dynamic' assessment of tumour cell proliferation in the clinical setting. Routine measurements can be achieved providing that well standardised tissue disaggregation and immunolabelling procedures are performed. In different solid tumours, cytokinetic results were satisfactory in 85% of cases as far as the BrdU-labelling index is concerned while 70% were acceptable for DNA synthesis time and tumour potential doubling time. Causes of failure are also considered and discussed. In order to guarantee the finest detection of aneuploidy, routine high resolution single-parameter DNA analysis is suggested. Our experience with further improvements in cell kinetic studies based on the possibility of gating the BrdU/DNA analysis for a subpopulation of interest (i.e. cytokeratin-positive cells in epithelial tumours) is also reported.
KW - DNA-BrdU-cytokeratin
KW - Flow cytometry
KW - Proliferation
KW - Solid tumours
UR - http://www.scopus.com/inward/record.url?scp=0029940130&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0029940130&partnerID=8YFLogxK
M3 - Article
VL - 10
SP - 101
EP - 113
JO - Analytical Cellular Pathology
JF - Analytical Cellular Pathology
SN - 0921-8912
IS - 2
ER -